Determination of a novel Aurora-A (AurA) kinase AKI603 by UPLC-MS/MS and its application to a bioavailability study in rat.

Abstract A simple, sensitive and accurate ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the determination of AKI603 in rat plasma has been firstly developed and validated. After a simple liquid–liquid extraction (LLE) with ethyl acetate, the analytes were separated on C18 column (2.1 × 100 mm, 1.9 μm, Thermo) by gradient elution with mobile phase of water (A) (containing 5 mM ammonium acetate and 0.1% formic acid) and methanol (B) with a flow rate of 0.3 mL min −1 and then analyzed by mass spectrometry in the positive multiple reactions monitoring (MRM) mode. The mass transitions monitored were m / z 410.0 → 352.9, m / z 457.1 → 367.9 for AKI603 and internal standard (Ly-7z), respectively. The developed method was validated for specificity, linearity and lower limit of quantification, intra- and inter-day precision and accuracy, extraction recovery, matrix effect and stability whose values satisfied the acceptable limits. The calibration curves for AKI603 was linear in concentration ranges of 0.025–5000 ng mL −1 . The method has been successfully used to the bioavailability study of AKI603 administered to rats intravenously (2.5 mg/kg) or orally (25 mg/kg). The oral bioavailability of AKI603 in rats was calculated as 28.7 ± 9.7%.